Intestinal trefoil factor binds to intestinal epithelial cells and induces nitric oxide production: Priming and enhancing effects of mucin

Xiaodi Tan*, Qian Ping Liu, Wei Hsueh, Yi-Hua Chen, Hong Chang, Frank Gonzalez-Crussi

*Corresponding author for this work

Research output: Contribution to journalArticle

25 Scopus citations

Abstract

Intestinal trefoil factor (ITF or TFF3), NO and epithelium-associated mucin have important roles in sustaining mucosal integrity in the gastrointestinal tract. In the present study we examined ITF-binding molecules on IEC-18 cells (an intestinal epithelial cell line) with the use of flow cytometry and localized these molecules on the cell surface by confocal microscopy. Furthermore, we studied the interaction of mucin and ITF and their co-operative effect on NO production by the epithelium. Stimulation of cells with mucin (5 mg/ml) for 90 min resulted in a 5-fold increase in ITF binding. Treatment of IEC-18 cells with actinomycin D or cycloheximide attenuated mucin-enhanced ITF binding. Ligand blot analysis confirmed the induction of ITF-binding protein in IEC-18 cells by mucin. These results indicate that transcriptional and translational mechanisms are involved in the effect of mucin. Treatment with ITF overnight resulted in a low level of nitrite production by the cells, a 5-fold increase over control, in a concentration-dependent manner. ITF-induced NO production was attenuated by 1400 W, a selective type II nitric oxide synthase (NOS2) inhibitor. By immunoblotting we found that NOS2 was up-regulated by ITF treatment. Priming IEC-18 cells with mucin for 90 min enhanced the effect of ITF on NO production, suggesting that the upregulation of ITF-binding molecules by mucin might be physiologically relevant. Taken together, these observations indicate (1) that ITF-binding molecules that are up-regulated by mucin exist on the intestinal epithelial surface, and (2) that ITF modulates epithelial NO production via the NOS2 pathway, which is enhanced by mucin.

Original languageEnglish (US)
Pages (from-to)745-751
Number of pages7
JournalBiochemical Journal
Volume338
Issue number3
DOIs
StatePublished - Mar 15 1999

Keywords

  • Confocal microscopy
  • Epithelial restitution
  • Flow cytometry
  • Intestinal-trefoil-factor-binding protein
  • Nitric oxide synthase 2

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Fingerprint Dive into the research topics of 'Intestinal trefoil factor binds to intestinal epithelial cells and induces nitric oxide production: Priming and enhancing effects of mucin'. Together they form a unique fingerprint.

  • Cite this